Meloidogyne incognita is one of the most polyphagous species of root-knot nematodes occurring in Brazil and worldwide. Eight M. incognita isolates were studied, representing two enzymatic phenotypes (esterase and malate desydrogenase: I1/N1, I2/N1) and four cryptic Meloidogyne sp.1 (S2/N1) isolates, representing one cytological type (3n040-46). Three M. hispanica isolates (Hi3/N1, 2n032-36) and two of an atypical Meloidogyne sp.2 (S2a/N3, 3n040-44) were included in this study for comparison. All isolates were tested with three M. incognita-specific molecular markers. The primer pairs B06F/R, miF/R and incK14F/R amplified three species-specific fragments of 1,200 bp, 955 bp and 399 bp, respectively for M. incognita and Meloidogyne sp.1 isolates. No amplification occurred in the M. hispanica and Meloidogyne sp.2 isolates, except with primers miF/R (1,650 bp). The genetic variability of the Meloidogyne spp. isolates was evaluated, using RAPD and ISSR markers. The phylogenetic analyses revealed two strongly supported monophyletic clades: clade I, consisting of M. hispanica and the atypical Meloidogyne sp.2 isolates, and clade II, clustering together all M. incognita and the Meloidogyne sp.1 isolates. Considering the biometrical, cytological and molecular approaches, it was possible to conclude that the isolates with three enzymatic phenotypes (I1/N1, I2/N1 and S2/N1) presented the characteristics described for M. incognita. Some correlations were detected between the isozymatic phenotypes and the tree topology (S2a/N3, Hi3/N1, I1/N1, S2/N1), but no strict correlation could be observed for the phenotype I2/N1 and one isolate of S2/N1. Morphologically, the Msp.2 isolates differ from M. incognita and M.

Accessions of Gossypium spp., some of them never previously tested, were evaluated for resistance to a local isolate of Meloidogyne incognita race 3 in greenhouse experiments. Nematode infection was characterized based on the galling and egg mass indexes and the reproduction factors (RF). Root-knot nematode reproduction among the newly tested accessions varied from highly susceptible (AS0188, AS0189) to moderately resistant (MT123 no. 3), and some accessions showed highly reduced nematode reproduction (CIR1343, CIR1348, Fai Mui). Histological observations of two resistant accessions (G. barbadense CIR1348 and G. hirsutum TX-25, respectively) showed that resistance occurs through a two-stage mechanism in the first accession and through a single-stage mechanism in the second. Parasitism is blocked early after second-stage juvenile (J2) penetration or during its initial tissue migration (CIR1348) and the development of later-stage juveniles into female adults is suppressed at a later stage (TX-25 and CIR1348). Fluorescence and bright light microscopy showed that root cells surrounding nematodes exhibit a hypersensitivity-like reaction, with the accumulation of presumably phenolic compounds and the presence of necrotic cells that limit the development of nematodes and the formation of giant cells. Underdeveloped giant cells with degenerated cytoplasmic content were found in small numbers in CIR1348 and in large numbers in TX-25, along with deformed nematodes. The full characterization of the defence mechanisms of novel sources of resistance to the root-knot nematode in cotton constitutes a first step towards their use in crop improvement.

Thirteen Meloidogyne arenaria isolates representing two cytological types (3n = 51-56, 2n = 42-48) and four enzymatic phenotypes (esterase and malate dehydrogenase: A3N1, A2N1, A1N1, A2N3) were studied using different approaches. The analysis of molecular markers showed a high level of polymorphism among the isolates. The trees obtained with RAPD or ISSR polymorphisms showed concordant results and agree with morphological studies. By considering morphometrical and morphological features, it was possible to conclude that the isolate with enzymatic phenotype A2N3 race 1 was the M. arenaria described in 1949 by Chitwood and appearing clearly separated in the trees, as well as in the outgroups. The seven isolates with phenotype A2N1 from different localities and isolate A1N1 can be considered morphometrically typical of M. arenaria race 2 and they were apparently clustered by geographical origin. Morphologically, they differed from isolate A2N3 race 1. The two isolates with phenotype A3N1 appeared to be closely related to the isolate of M. morocciensis and, considering all of the features described for this species, were identified as such. The two isolates A2N3 race 2 were identified either as an atypical M. arenaria or an unidentified species (females and males having atypical stylets), and clustered together and separated from other M. arenaria isolates with high bootstrap support. The same M. arenaria isolates were tested with the species-specific molecular marker, type SCAR. A fragment of 420 bp was obtained for ten isolates of M. arenaria, including the atypical A2N3 race 2 and M. morocciensis. This fragment was not amplified for three typical A2N1 isolates of M. arenaria.

An obligate parasite bacterium of the root-knot nematode, Pasteuria penetrans strain P10, isolated from Meloidogyne incognita females on banana roots in Imperatriz Maranhão State, Brazil, was evaluated in glasshouse conditions, using two doses of a dry root bionematicide (10 7 endospores (5.0 g/seedling) and 10 6 endospores (0.5 g/seedling)) on seedlings of cv. Mundo Novo coffee. The soil in which coffee seedlings were raised was inoculated previously with these two doses of P. penetrans and after 2 months the plants were transferred to soils of different textures: clay-sandy soil (38% clay, 2% silt and 60% sand) and sandy soil (17% clay, 0% silt and 83% sand). When the coffee plants were 30 cm high, they were inoculated with 20 000 eggs/plant of M. incognita race 1. The coffee plants were examined 8, 16 and 24 months after nematode plant infestation. The effectiveness of the biological control was determined by the reduction of nematode reproduction factor, which ranged from 62 to 67% in clay-sandy soil and 80 to 85% in sandy soil. The mechanism of suppression caused by the bacterium was evaluated by the percentage of infected second-stage juveniles (J2), number of endospores attached/J2 and number of infected females. The high levels of suppression were related to time, increasing from 8 to 24 months, and to the percentage of sand in the soil.